Seven groups of biomedical NMR researchers at the University of Pennsylvania propose to carry out high-field, high-resolution, multi-nuclear NMR studies by modernizing the existing Bruker WH-360 NMR spectrometer system in the Medical School NMR Facility. The proposed research projects deal with (1) high-resolution, high-field proton NMR studies of the subunit cooperativity and allostery of hemoglobin using cobalt-substituted hemoglobins, (2) phosphorus NMR examination of thermodynamic performance of mitochondrial suspension, (3) phosphorus NMR study of thermodynamics of sodium transport and intracellular pHs in frog skin, (4) cadmium, phosphorus, and proton NMR studies of active site characterization of yeast inorganic pyrophosphatase, (5) fluorine NMR studies of transmembrane pH/electrical potential gradients during changes in the metabolic or developmental state of the tissue, cell, or organelle, (6) single-cell phosphorus NMR studies of phosphometabolite profiles using giant muscle cells from the barnacle, Balanus nubilis, and (7) proton, phosphorus, and carbon NMR studies of metabolic states in heart, liver, and brain tissues.